Effects of Secretome from Dynamic 3D Cell Culture System onto Growth and Cytoprotection of Nasal Fibroblast

IF 0.1 Q4 MEDICINE, GENERAL & INTERNAL
Siti Ni, K. Kavireshna, Zafirah Mr, Y. W. Foo, R. Rabiatuladawiyah, S. Husain, A. Mohd-Ilham, Y. Lokanathan
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引用次数: 1

Abstract

Three dimensional (3D) models mimic the features of native tissue environment. Thus, morphology and signalling of cells from 3D culture are often more physiological than routine two dimensional (2D) cell culture. It is also known that the cell-secreted products have paracrine effect on other cells growth. In this experimental study, we optimised the nasal fibroblast culture on a 3D cell culture system and studied the effects of secretome from the 3D culture (3DCM) onto fibroblast growth and cytoprotection. Nasal fibroblast was isolated from human nasal turbinates. The suitable microcarrier was selected by culturing the fibroblasts in passage 3 on various types PolyGEM™ polystyrene microcarriers. Then, the cells were cultured on selected microcarrier using a 3D culture system and the conditioned medium (CM) was collected. 3DCM were supplemented to fibroblasts to study for attachment, proliferation, and cytoprotective effects against cytotoxicity of Centella asiatica. Bicinchonic Acid Assay (BCA) was performed to quantify protein amount in CMs. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) was performed for preliminary profiling and comparison of 2DCM and 3DCM protein profile. Our study showed that the 3DCM did not significantly enhance cell attachment and proliferation. The secretome of both 2DCM and 3DCM found to have significant cytoprotective effect onto nasal fibroblast against cytotoxicity of C.asiatica extract. 3DCM had higher protein concentration than 2DCM. SDS-PAGE showed three exclusive proteins in 3DCM and four exclusive proteins in 2DCM. Future study should be conducted on utility of nasal fibroblast secretome on cytoprotection against harmful agents in environment and cytotoxicity of natural products.
动态三维细胞培养系统分泌组对鼻成纤维细胞生长和细胞保护的影响
三维(3D)模型模拟了原生组织环境的特征。因此,与常规的二维(2D)细胞培养相比,3D培养细胞的形态和信号传导通常更具生理性。我们也知道,细胞分泌的产物对其他细胞的生长有旁分泌作用。在本实验研究中,我们优化了鼻腔成纤维细胞在3D细胞培养系统上的培养,并研究了3D培养(3DCM)分泌组对成纤维细胞生长和细胞保护的影响。从人鼻鼻甲中分离到鼻成纤维细胞。将第3代成纤维细胞培养在不同类型PolyGEM™聚苯乙烯微载体上,选择合适的微载体。然后用三维培养系统在选定的微载体上培养细胞,收集条件培养基(CM)。将3DCM添加到成纤维细胞中,研究积雪草的附着、增殖和细胞保护作用。采用Bicinchonic Acid Assay (BCA)测定CMs中的蛋白含量。采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)对2DCM和3DCM进行初步分析和比较。我们的研究表明,3DCM对细胞的附着和增殖没有明显的促进作用。2DCM和3DCM的分泌组对鼻部成纤维细胞均有明显的细胞保护作用。3DCM的蛋白浓度高于2DCM。SDS-PAGE显示3DCM中有3个特异性蛋白,2DCM中有4个特异性蛋白。鼻腔成纤维细胞分泌组对环境中有害物质的细胞保护作用和天然产物的细胞毒性作用有待进一步研究。
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来源期刊
Medicine and Health
Medicine and Health MEDICINE, GENERAL & INTERNAL-
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